Abstract: 【Objective】To understand the protoplasts isolation and regeneration of different edible （medicine） fungi, so as to provide materials for protoplast mutagenesis and fusion, so as to selectively breed new strains with outstanding excellent characters.【Methods】Five kinds of edible （medicine） fungi mycelia were used as test materials, and five kinds of edible （medicine） fungi protoplasts were prepared by enzymatic hydrolysis.【Results】under the same experimental conditions,the amount of protoplast released: Sanghuangporus zsh> Grassland white mushroom BM1716> Pleurotus eryngii xb> Grassland black mushroom hm8124-1 > Hypsizygus marmoreus xwg; the regeneration rate：Hypsizygus marmoreus xwg > Pleurotus eryngii xb> Grassland blacmushroom hm8124-1> Grassland white mushroom BM1716>Sanghuangporus zsh.It was observed that the single colony of protoplast regeneration of Grassland black mushroom hm8124-1,thick and dense,while the single colony of protoplast regeneration of 6 Grassland white mushroom BM1716 were significantly different,the growth morphology and growth speed of the colony were different,and the phenomenon of angular change appeared.【Conclusion】The release and regeneration rate of protoplasts of different edible （medicine） fungi are quite different.It is necessary to optimize the regeneration medium in subsequent experiments. The growth of regeneration colony of single protoplast of 6 Grassland white mushroom BM1716 were observed,and the difference was very obvious. It is possible that somaclonal variation occurred in the process of protoplast isolation and regeneration.

Key words: Edible（medicinal）fungi, Protoplast separation, Protoplast regeneration